Forms of a PI3K delta selective inhibitor for use in pharmaceutical formulations

ABSTRACT

The present invention relates to solid state forms of a p-toluenesulfonic acid salt (PTSA) of the selective PI3K delta inhibitor (S)-2-(1-(4-amino-3-(3-fluoro-4-isopropoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-6-fluoro-3-(3-fluorophenyl)-4H-chromen-4-one (TGR-1202). The present invention also relates to methods of preparing the same, pharmaceutical compositions containing them, and methods of treating a PI3K kinase mediated disease or disorder, such as cancer, by administering the same.

This application is a continuation of U.S. patent application Ser. No.15/950,606, filed Apr. 11, 2018, which is a divisional of U.S. patentapplication Ser. No. 15/313,454, filed Nov. 22, 2016, now U.S. Pat. No.9,969,740, which is the U.S. national stage of International PatentApplication No. PCT/IB2015/053940, filed May 26, 2015, which claims thebenefit of Indian Patent Application Nos. 2596/CHE/2014, filed May 27,2014, and 2597/CHE/2014, filed May 27, 2014, each of which is herebyincorporated by reference in its entirety.

FIELD OF THE INVENTION

The present invention relates to solid state forms of ap-toluenesulfonic acid salt (PTSA) of the selective PI3K delta inhibitor(S)-2-(1-(4-amino-3-(3-fluoro-4-isopropoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-6-fluoro-3-(3-fluorophenyl)-4H-chromen-4-one(TGR-1202). The present invention also relates to methods of preparingthe same, pharmaceutical compositions containing them, and methods oftreating a PI3K kinase mediated disease or disorder, such as cancer, byadministering the same.

BACKGROUND OF THE INVENTION

TGR-1202, chemically known as(S)-2-(1-(4-amino-3-(3-fluoro-4-isopropoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-6-fluoro-3-(3-fluorophenyl)-4H-chromen-4-one,has the following chemical structure:

The preparation of TGR-1202 and its salts is described in InternationalPublication No. WO 2014/006572 and U.S. Patent Publication No.2014/0011819, each of which is incorporated herein by reference in itsentirety for all purposes. TGR-1202 is an investigational drug currentlyundergoing multiple clinical trials in the area of haematologicalmalignancies.

WO 2014/006572 and US 2014/0011819 describe the synthesis of TGR-1202(Example B1) and also disclose the therapeutic activity of this moleculeto inhibit, regulate and/or modulate the signal transduction of PI3K.

SUMMARY OF THE INVENTION

The present invention relates to new solid state forms of ap-toluenesulfonic acid salt (PTSA) of the selective PI3K delta inhibitor(S)-2-(1-(4-amino-3-(3-fluoro-4-isopropoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-6-fluoro-3-(3-fluorophenyl)-4H-chromen-4-one(TGR-1202) (e.g.,(S)-2-(1-(4-amino-3-(3-fluoro-4-isopropoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-6-fluoro-3-(3-fluorophenyl)-4H-chromen-4-one4-methyl-benzenesulfonate).

The present inventors have discovered that particles of the PTSA salt ofTGR-1202 having certain particle sizes exhibit enhanced solubility andpharmacokinetics upon oral administration.

In one embodiment, the PTSA salt has a d(0.9) of from about 5 to about50 μm, such as from about 5 to about 25 μm or from about 5 to about 15μm. The PTSA salt may also have a d(0.5) of from about 1 to about 10 μm,such as from about 2.0 to about 10 μm, from about 1 to about 5 μm, orfrom about 2.0 to about 5 μm. The PTSA salt may have a d(0.1) of fromabout 0.5 to about 1.5 μm, such as from about 0.5 to about 1.0 μm.

In another embodiment, the PTSA salt of TGR-1202 has a d(0.5) of fromabout 1 to about 10 μm, such as from about 2.0 to about 10 μm, fromabout 1 to about 5 μm, or from about 2.0 to about 5 μm. The PTSA saltmay also have a d(0.1) of from about 0.5 to about 1.5 μm, such as fromabout 0.5 to about 1.0 μm. The PTSA salt may have a d(0.9) of from about5 to about 50 μm, such as from about 5 to about 25 μm or from about 5 toabout 15 μm.

In yet another embodiment, the PTSA salt has a d(0.1) of from about 0.5to about 1.5 μm, such as from about 0.5 to about 1.0 μm. The PTSA saltmay also have a d(0.9) of from about 5 to about 50 μm, such as fromabout 5 to about 25 μm or from about 5 to about 15 μm. The PTSA salt mayhave a d(0.9) of from about 5 to about 50 μm, such as from about 5 toabout 25 μm or from about 5 to about 15 μm.

In another embodiment, the PTSA salt of TGR-1202 has a particle sizedistribution (PSD) where

-   -   (i) the d(0.1) is from about 0.5 to about 1.5 μm, d(0.5) is from        about 2.0 to about 10 μm and d(0.9) is from about 5.0 to about        50 μm;    -   (ii) the d(0.1) is from about 0.5 to about 1.5 μm, d(0.5) is        from about 2.0 to about 5.0 μm and d(0.9) is from about 5.0 to        about 50 μm;    -   (iii) the d(0.1) is from about 0.5 to about 1.5 μm, d(0.5) is        from about 2.0 to about 10 μm and d(0.9) is from about 5.0 to        about 25 μm; or    -   (iv) the d(0.1) is from about 0.5 to about 1.0 μm, d(0.5) is        from about 2.0 to about 5 μm and d(0.9) is from about 5.0 to        about 15 μm.

In yet another embodiment, the PTSA salt of TGR-1202 has a particle sizedistribution (PSD) where the d(0.1) is from about 0.5 to about 1.5 μm,d(0.5) is from about 2.0 to about 10 μm and d(0.9) is from about 5.0 toabout 50 μm.

In yet another embodiment, the PTSA salt of TGR-1202 has a particle sizedistribution (PSD) where the d(0.1) is from about 0.5 to about 1.5 μm,d(0.5) is from about 2.0 to about 5.0 μm and d(0.9) is from about 5.0 toabout 50 μm.

In yet another embodiment, the PTSA salt of TGR-1202 has a particle sizedistribution (PSD) where the d(0.1) is from about 0.5 to about 1.0 μm,d(0.5) is from about 2.0 to about 10.0 μm and d(0.9) is from about 5.0to about 15 μm.

In yet another embodiment, the PTSA salt of TGR-1202 has a particle sizedistribution (PSD) where the d(0.1) is from about 0.5 to about 1.0 μm,d(0.5) is from about 2.0 to about 5.0 μm and d(0.9) is from about 5.0 toabout 15 μm.

In yet another embodiment, at most 5%, at most 3%, at most 2%, or atmost 1% of the particles of the PTSA salt of TGR-1202 have a particlesize less than 1.0 microns. At the same time, at least 95%, at least96%, at least 97%, at least 98%, at least 99%, or at least 99.5% of theparticles of the PTSA salt of TGR-1202 have a particle size less than100 microns. In one particular embodiment, the PTSA salt of TGR-1202also has a d(0.1) of from about 0.5 to about 1.5 μm, d(0.5) of fromabout 2.0 to about 10 μm and d(0.9) of from about 5.0 to about 50 μm. Inanother embodiment, the PTSA salt of TGR-1202 also has a d(0.1) of fromabout 0.5 to about 1.5 μm, d(0.5) of from about 2.0 to about 5.0 μm andd(0.9) of from about 5.0 to about 50 μm. In yet another embodiment, thePTSA salt of TGR-1202 also has a d(0.1) of from about 0.5 to about 1.5μm, d(0.5) of from about 2.0 to about 10 μm and d(0.9) of from about 5.0to about 25 μm. In yet another embodiment, the PTSA salt of TGR-1202 hasa d(0.1) of from about 0.5 to about 1.0 μm, d(0.5) of from about 2.0 toabout 5.0 μm and d(0.9) of from about 5.0 to about 15 μm.

In yet another embodiment, at most 5%, at most 3%, at most 2%, or atmost 1% of the particles of the PTSA salt of TGR-1202 have a particlesize less than 1.0 microns. At the same time, at least 95%, at least96%, at least 97%, at least 98%, at least 99%, or at least 99.5% of theparticles of the PTSA salt of TGR-1202 have a particle size less than 50microns. In one particular embodiment, the PTSA salt of TGR-1202 alsohas a d(0.1) of from about 0.5 to about 1.5 μm, d(0.5) of from about 2.0to about 10 μm and d(0.9) of from about 5.0 to about 50 μm. In anotherembodiment, the PTSA salt of TGR-1202 also has a d(0.1) of from about0.5 to about 1.5 μm, d(0.5) of from about 2.0 to about 5.0 μm and d(0.9)of from about 5.0 to about 50 μm. In yet another embodiment, the PTSAsalt of TGR-1202 also has a d(0.1) of from about 0.5 to about 1.5 μm,d(0.5) of from about 2.0 to about 10 μm and d(0.9) of from about 5.0 toabout 25 μm. In yet another embodiment, the PTSA salt of TGR-1202 has ad(0.1) of from about 0.5 to about 1.0 μm, d(0.5) of from about 2.0 toabout 5.0 μm and d(0.9) of from about 5.0 to about 15 μm.

In yet another embodiment, at most 5%, at most 3%, at most 2%, or atmost 1% of the particles of the PTSA salt of TGR-1202 have a particlesize less than 1.0 microns. At the same time, at least 95%, at least96%, at least 97%, at least 98%, at least 99%, or at least 99.5% of theparticles of the PTSA salt of TGR-1202 have a particle size less than 25microns. In one particular embodiment, the PTSA salt of TGR-1202 alsohas a d(0.1) of from about 0.5 to about 1.5 μm, d(0.5) of from about 2.0to about 10 μm and d(0.9) of from about 5.0 to about 25 μm. In yetanother embodiment, the PTSA salt of TGR-1202 has a d(0.1) of from about0.5 to about 1.0 μm, d(0.5) of from about 2.0 to about 5.0 μm and d(0.9)of from about 5.0 to about 15 μm.

In yet another embodiment, at most 5%, at most 3%, at most 2%, or atmost 1% of the particles of the PTSA salt of TGR-1202 have a particlesize less than 1.0 microns. At the same time, at least 95%, at least96%, at least 97%, at least 98%, at least 99%, or at least 99.5% of theparticles of the PTSA salt of TGR-1202 have a particle size less than 15microns. In one particular embodiment, the PTSA salt of TGR-1202 alsohas a d(0.1) of from about 0.5 to about 1.5 μm, d(0.5) of from about 2.0to about 10 μm and d(0.9) of from about 5.0 to about 25 μm. In yetanother embodiment, the PTSA salt of TGR-1202 has a d(0.1) of from about0.5 to about 1.0 μm, d(0.5) of from about 2.0 to about 5.0 μm and d(0.9)of from about 5.0 to about 15 μm.

In yet another embodiment, the PTSA salt of TGR-1202 exhibits an X-raypowder diffraction (XRPD) pattern substantially as shown in FIG. 1(hereinafter referred to as Form A).

In yet another embodiment, the PTSA salt of TGR-1202 exhibits adifferential scanning calorimeter (DSC) pattern having a characteristicendothermic peak at about 154° C. (Form A).

In yet another embodiment, the PTSA salt of TGR-1202 exhibits an XRPDpattern exhibiting one or more peaks selected from 5.0, 10.1, 15.9,16.1, 16.3, 20.0, 22.1, and 24.4±0.05, 0.1, or 0.2° 2Θ. For example, thesalt may exhibit an XRPD pattern having one or more peaks (such as 2, 3,4, 5, 6, 7 or 8 peaks) selected from 5.01, 10.09, 15.91, 16.13, 16.34,20.00, 22.06 and 24.42±0.05, 0.1, or 0.2° 2Θ.

In yet another embodiment, the PTSA salt of TGR-1202 exhibits an XRPDpattern substantially as shown in FIG. 2 (hereinafter referred to asForm B).

In yet another embodiment, the PTSA salt of TGR-1202 exhibits adifferential scanning calorimeter (DSC) pattern having a characteristicendothermic peak at about 146° C. (Form B).

In yet another embodiment, the PTSA salt of TGR-1202 exhibits an XRPDpattern having one or more peaks selected from 5.0, 10.1, 22.1, and24.5±0.05, 0.1, or 0.2° 2θ. For example, the salt may exhibit an XRPDpattern having one or more peaks selected from 5.04, 10.13, 22.11 and24.45±0.05, 0.1, or 0.2° 2θ.

Yet another embodiment is crystalline Form B of a PTSA salt of TGR-1202having a particle size distribution (PSD) which is defined by a d(0.1)of from about 0.5 to about 1.5 μm, d(0.5) of from about 2.0 to 10 μm andd(0.9) of from about 5 to about 50 μm.

Yet another embodiment is crystalline Form B of a PTSA salt of TGR-1202having a particle size distribution (PSD) which is defined by a d(0.1)of from about 0.5 to about 1.5 μm, d(0.5) of from about 2.0 to about 5μm and d(0.9) of from about 5 to about 25 μm.

Yet another embodiment is crystalline Form B of a PTSA salt of TGR-1202having a particle size distribution (PSD) which is defined by a d(0.1)of from about 0.5 to about 1.0 μm, d(0.5) of from about 2.0 to about 5μm and d(0.9) of from about 5 to about 15 μm.

The present invention further provides a pharmaceutical compositioncomprising a PTSA salt of TGR-1202 according to any of the embodimentsdescribed herein (e.g., having a particle size distribution as definedby any embodiment herein) and a pharmaceutically acceptable excipient.

The present invention further provides a pharmaceutical compositioncomprising crystalline Form B of a PTSA salt of TGR-1202 as described inany embodiment herein and a pharmaceutically acceptable excipient.

The present invention further provides a pharmaceutical compositioncomprising crystalline Form B of a PTSA salt of TGR-1202 having aparticle size distribution as defined in any embodiment herein and apharmaceutically acceptable excipient.

Another embodiment is a PTSA salt of TGR-1202 having an XRPD patternsubstantially as shown in FIG. 2, suitable for use in a pharmaceuticalcomposition for the treatment of a PI3K associated disease, disorder orcondition, e.g., a proliferative disease such as cancer.

Another embodiment is a PTSA salt of TGR-1202 having a particle sizedistribution as defined in any embodiment herein, suitable for use in apharmaceutical composition for the treatment of a PI3K associateddisease, disorder or condition, e.g., a proliferative disease such ascancer.

The present invention further provides a pharmaceutical compositioncomprising crystalline Form B of a PTSA salt of TGR-1202 having aparticle size distribution as defined in any embodiment herein and apharmaceutically acceptable excipient. The pharmaceutical compositionmay further comprise one or more of additional active ingredients, suchas other active agents (such as anti-cancer agents and the active agentsdiscussed below).

The invention further provides a pharmaceutical composition comprising aPTSA salt of TGR-1202 having a particle size distribution as defined inany embodiment herein and a pharmaceutically acceptable excipient. Thepharmaceutical composition may further comprise one or more ofadditional active ingredients, such as other active agents (such asanti-cancer agents and the active agents discussed below).

One embodiment is a solid oral pharmaceutical composition (such as atablet) comprising a PTSA salt of TGR-1202 as described herein and oneor more solubilizers (such as hydroxypropyl betadex). The weight ratioof PTSA salt to solubilizer (such as hydroxypropyl betadex) can rangefrom about 1.5:1 to about 1:1.5. The composition may further include oneor more excipients selected from suspending agents (such ashydroxypropyl cellulose), dispersing agents (such as microcrystallinecellulose), disintegrants (such as croscarmellose sodium), lubricants(such as magnesium stearate), and any combination of any of theforegoing.

In additional embodiments, the PTSA salt as defined in any embodimentherein has an enantiomeric excess (e.e.) of at least about 60%, at leastabout 75%, at least about 80%, at least about 85%, at least about 90%,at least about 95%, at least about 98%, at least about 99%, at leastabout 99.5% or at least about 99.9%.

Yet another embodiment is a crystalline PTSA salt of TGR-1202, wherein(i) the crystalline salt exhibits an XRPD pattern having one or morepeaks (such as 2, 3 or 4 peaks) selected from 5.0, 10.1, 22.1, and24.5±0.2° 2Θ and (ii) the crystalline salt is substantially free ofother solid state forms of the PTSA salt.

Yet another embodiment is a crystalline PTSA salt of TGR-1202, wherein(i) the crystalline salt exhibits an XRPD pattern having one or morepeaks (such as 2, 3 or 4 peaks) selected from 5.0, 10.1, 22.1, and24.5±0.2° 2Θ and (ii) the crystalline salt contains less than 5% (suchas less than 4%, less than 3%, less than 2% or less than 1%) of othersolid state forms of the PTSA salt.

In another aspect, the present invention relates to a method ofpreparing a crystalline PTSA salt of TGR-1202 (i.e., a p-toluenesulfonicacid salt of(S)-2-(1-(4-amino-3-(3-fluoro-4-isopropoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-6-fluoro-3-(3-fluorophenyl)-4H-chromen-4-one).In one embodiment, the method comprises removing the solvent from amixture of a PTSA salt of TGR-1202 and an ether solvent, such asdi-tert-butyl ether, diethyl ether, diethylene glycol diethyl ether,diisopropyl ether, dimethoxyethane, dimethoxymethane, ethyl tert-butylether, methoxyethane, 2-(2-methoxyethoxy)ethanol, methyl tert-butylether, 2-methyltetrahydrofuran, tetrahydrofuran and tetrahydropyran. Themixture is preferably stirred (for example, for 3, 6, 12, or more hours)prior to removing the solvent. The solvent may be removed by drying. Inone preferred embodiment, the method comprises stirring, filtering anddrying a mixture of a PTSA salt of TGR-1202 and an ether solvent. Themixture is preferably a suspension, which can, for example, be formed bysuspending a PTSA salt of TGR-1202 in an ether solvent.

In another aspect, the present invention is directed to a method ofinhibiting PI3K delta in a patient in need thereof by administering tothe patient an effective amount of a PTSA salt of TGR-1202 as describedherein (e.g., crystalline Form B of the PTSA salt of TGR-1202) andhaving a particle size distribution as defined in any embodimentherein).

Yet another embodiment is a method of treating, preventing, and/orinhibiting a PI3K protein kinase mediated disease, disorder or condition(such as cancer or other proliferative disease or disorder) in a patientby administering to the patient an effective amount of a PTSA salt ofTGR-1202 as described herein (e.g., crystalline Form B of the PTSA saltof TGR-1202).

Yet another embodiment is a method of treating a PI3K associateddisease, disorder or condition in a patient by administering to thepatient an effective amount of a PTSA salt of TGR-1202 as describedherein (e.g., crystalline Form B of the PTSA salt of TGR-1202). In oneembodiment, the amount of the crystalline form B of the PTSA salt ofTGR-1202 administered is sufficient to treat a PI3K associated disease,disorder or condition by inhibition of PI3K delta.

Yet another embodiment of the present invention is a method for treatinga proliferative disease by administering to a patient in need of suchtreatment an effective amount of a PTSA salt of TGR-1202 as describedherein (e.g., crystalline Form B of the PTSA salt of TGR-1202). In oneembodiment, the amount of crystalline form B of the PTSA salt ofTGR-1202 administered is sufficient to treat the proliferative diseaseby inhibition of PI3K delta.

Yet another embodiment of the present invention is a method for treatinga proliferative disease by administering to a patient in need of suchtreatment an effective amount of a PTSA salt of TGR-1202 as describedherein (e.g., crystalline Form B of the PTSA salt of TGR-1202), incombination (simultaneously or sequentially) with at least one otheranti-cancer agent. In one embodiment, the amount of PTSA salt ofTGR-1202 (e.g., Form B of the PTSA salt of TGR-1202) administered issufficient to treat (or facilitate treatment of) the proliferativedisease by inhibition of PI3K delta.

Yet another embodiment is a method of treating a PI3K associateddisease, disorder or condition in a patient, comprising administering toa patient in need of such treatment an effective amount of a PTSA saltof TGR-1202 as described herein (e.g., crystalline Form B of PTSA saltof TGR-1202), optionally admixed with at least one pharmaceuticallyacceptable excipients. In particular embodiments, the compositioncomprises a therapeutically effective amount of crystalline Form B ofthe PTSA salt of TGR-1202 according to any embodiment herein (e.g., acrystalline Form B of the PTSA salt of TGR-1202 having a particle sizedistribution as defined in any embodiment herein) for the treatment of aPI3K associated disease, disorder or condition.

Specific embodiments provide a method of treating cancer in a patient,comprising administering to the patient a pharmaceutical compositioncomprising a PTSA salt of TGR-1202 as described herein (e.g.,crystalline Form B of PTSA salt of TGR-1202), optionally admixed with atleast one pharmaceutically acceptable excipient. In particularembodiments, the composition comprises of a therapeutically effectiveamount of crystalline Form B of PTSA salt of TGR-1202 according to anyembodiment herein (e.g., a crystalline Form B of PTSA salt of TGR-1202having a particle size distribution as defined in any embodiment herein)for the treatment of cancer in a patient.

The crystalline Form B of PTSA salt of TGR-1202 according to anyembodiment herein (e.g., a crystalline Form B of PTSA salt of TGR-1202having a particle size distribution as defined in any embodiment herein)is useful in the treatment of a variety of cancers, including, but notlimited to, the following:

-   -   carcinoma, including that of the bladder, breast, colon, kidney,        liver, lung (including small cell lung cancer), oesophagus, gall        bladder, uterus, ovary, testes, larynx, oral cavity,        gastrointestinal tract (e.g., oesophagus, stomach, pancreas),        brain, cervix, thyroid, prostate, blood, and skin (including        squamous cell carcinoma);    -   hematopoietic tumors of lymphoid lineage, including leukemia,        acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell        lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkins        lymphoma, hairy cell lymphoma and Burkett's lymphoma;    -   hematopoietic tumors of myeloid lineage, including acute and        chronic myelogenous leukemias, myelodysplastic syndrome and        promyelocytic leukemia;    -   tumors of mesenchymal origin, including fibrosarcoma and        rhabdomyosarcoma;    -   tumors of the central and peripheral nervous system, including        astrocytoma, neuroblastoma, glioma and schwannomas; and    -   other tumors, including melanoma, seminoma, teratocarcinoma,        osteosarcoma, xenoderoma pigmentosum, keratoctanthoma, thyroid        follicular cancer and Kaposi's sarcoma.

The PTSA salt of TGR-1202 described herein (including the crystallineForm B) as modulators of apoptosis are useful in the treatment,prevention, and inhibition of cancer (including, but not limited to,those types mentioned herein).

The PTSA salt of TGR-1202 described herein (including the crystallineForm B) are useful in the chemoprevention of cancer. Chemopreventioninvolves inhibiting the development of invasive cancer by blocking theinitiating mutagenic event, blocking the progression of pre-malignantcells that have already suffered an insult, or inhibiting tumor relapse.The PTSA salt, including Form B of it, is also useful in inhibitingtumor angiogenesis and metastasis. One embodiment of the invention is amethod of inhibiting tumor angiogenesis or metastasis in a patient byadministering an effective amount of a PTSA salt of TGR-1202 (such ascrystalline Form B of PTSA salt of TGR-1202).

Another embodiment of the present invention is a method of treating animmune system-related disease (e.g., an autoimmune disease), a diseaseor disorder involving inflammation (e.g., asthma, chronic obstructivepulmonary disease, rheumatoid arthritis, inflammatory bowel disease,glomerulonephritis, neuroinflammatory diseases, multiple sclerosis,uveitis and disorders of the immune system), cancer or otherproliferative disease, a hepatic disease or disorder, or a renal diseaseor disorder. The method includes administering an effective amount ofone or more compounds of the present invention.

Examples of immune disorders which can be treated by the compounds ofthe present invention include, but are not limited to, psoriasis,rheumatoid arthritis, vasculitis, inflammatory bowel disease,dermatitis, osteoarthritis, asthma, inflammatory muscle disease,allergic rhinitis, vaginitis, interstitial cystitis, scleroderma,osteoporosis, eczema, allogeneic or xenogeneic transplantation (organ,bone marrow, stem cells and other cells and tissues) graft rejection,graft-versus-host disease, lupus erythematosus, inflammatory disease,type I diabetes, idiopathic pulmonary fibrosis (IPF) (or cryptogenicfibrosing alveolitis (CFA) or idiopathic fibrosing interstitialpneumonia), pulmonary fibrosis, dermatomyositis, Sjogren's syndrome,thyroiditis (e.g., Hashimoto's and autoimmune thyroiditis), myastheniagravis, autoimmune hemolytic anemia, multiple sclerosis, cysticfibrosis, chronic relapsing hepatitis, primary biliary cirrhosis,allergic conjunctivitis and atopic dermatitis.

Yet another embodiment is a method of treating leukemia in a patient byadministering a therapeutically effective amount of a compound of thepresent invention. For example, the compounds of the present inventionare effective for treating chronic lymphocytic leukemia (CLL),non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), multiplemyeloma (MM), small lymphocytic lymphoma (SLL), and indolentnon-Hodgkin's lymphoma (I-NHL).

In the aforementioned methods of treatment, one or more additionalactive agents can be administered with a PTSA salt of TGR-1202, such asthe crystalline form B of PTSA salt of TGR-1202 as described in anyembodiment herein. For example, the PTSA salt of TGR-1202 as describedin any embodiment herein is useful in combination (administered togetheror sequentially) with known anti-cancer treatments such as chemotherapy,radiation therapy, biological therapy, bone marrow transplantation, stemcell transplant or any other anticancer therapy or with one or morecytostatic, cytotoxic or anticancer agents or targeted therapy eitheralone or in combination, such as but not limited to, for example, DNAinteractive agents, such as fludarabine, cisplatin, chlorambucil,bendamustine or doxorubicin; alkylating agents, such ascyclophosphamide; topoisomerase II inhibitors, such as etoposide;topoisomerase I inhibitors, such as CPT-11 or topotecan; tubulininteracting agents, such as paclitaxel, docetaxel or the epothilones(for example ixabepilone), either naturally occurring or synthetic;hormonal agents, such as tamoxifen; thymidilate synthase inhibitors,such as 5-fluorouracil; anti-metabolites, such as methotrexate; othertyrosine kinase inhibitors such as Iressa and OSI-774; angiogenesisinhibitors; EGF inhibitors; VEGF inhibitors; CDK inhibitors; SRCinhibitors; c-Kit inhibitors; Her1/2 inhibitors and monoclonalantibodies directed against growth factor receptors such as erbitux(EGF) and herceptin (Her2); CD20 monoclonal antibodies such asrituximab, ublixtumab (TGR-1101), ofatumumab (HuMax; Intracel),ocrelizumab, veltuzumab, GA101 (obinutuzumab), AME-133v (LY2469298,Applied Molecular Evolution), ocaratuzumab (Mentrik Biotech), PRO131921,tositumomab, hA20 (Immunomedics, Inc.), ibritumomab-tiuxetan, BLX-301(Biolex Therapeutics), Reditux (Dr. Reddy's Laboratories), and PRO70769(described in WO2004/056312); other B-cell targeting monoclonalantibodies such as belimumab, atacicept or fusion proteins such asblisibimod and BR3-Fc; other monoclonal antibodies such as alemtuzumab;CHOP (cyclophosphamide, doxorubicin, vincristine, prednisone); R-CHOP(rituximab-CHOP); hyperCV AD (hyperfractionated cyclophosphamide,vincristine, doxorubicin, dexamethasone, methotrexate, cytarabine);R-hyperCV AD (rituximab-hyperCV AD); FCM (fludarabine, cyclophosphamide,mitoxantrone); R-FCM (rituximab, fludarabine, cyclophosphamide,mitoxantrone); bortezomib and rituximab; temsirolimus and rituximab;temsirolimus and Velcade®; Iodine-131 tositumomab (Bexxar®) and CHOP-CVP(cyclophosphamide, vincristine, prednisone); R-CVP (rituximab-CVP); ICE(iphosphamide, carboplatin, etoposide); R-ICE (rituximab-ICE); FCR(fludarabine, cyclophosphamide, rituximab); FR (fludarabine, rituximab);and D.T. PACE (dexamethasone, thalidomide, cisplatin, adriamycin,cyclophosphamide, etoposide); and other protein kinase modulators.

The crystalline form B of PTSA salt of TGR-1202 as described in anyembodiment herein is also useful in combination (administered togetheror sequentially) with one or more steroidal anti-inflammatory drugs,non-steroidal anti-inflammatory drugs (NSAIDs) or immune selectiveanti-inflammatory derivatives (ImSAIDs).

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is an XRPD spectrum for the PTSA salt of TGR-1202 prepared by theprocess described in Example 1 (Form A).

FIG. 2 is an XRPD spectrum for the PTSA salt of TGR-1202 prepared by theprocess described in Example 2 (Form B).

FIG. 3 is a DSC trace for the PTSA salt of TGR-1202 prepared by theprocess described in Example 1 (Form A).

FIG. 4 is a DSC trace for the PTSA salt of TGR-1202 prepared by theprocess described in Example 2 (Form B).

DETAILED DESCRIPTION OF THE INVENTION

As used herein the following definitions shall apply unless otherwiseindicated.

When ranges are used herein for physical properties, such as molecularweight, or chemical properties, such as chemical formulae, allcombinations and subcombinations of ranges and specific embodimentstherein are intended to be included. The term “about” when referring toa number or a numerical range means that the number or numerical rangereferred to is an approximation within experimental variability (orwithin statistical experimental error), and thus the number or numericalrange may vary from, for example, between 1% and 15% of the statednumber or numerical range. The term “comprising” (and related terms suchas “comprise” or “comprises” or “having” or “including”) includes thoseembodiments, for example, an embodiment of any composition of matter,composition, method, or process, or the like, that “consist of” or“consist essentially of” the described features.

The following abbreviations and terms have the indicated meaningsthroughout: PI3-K=Phosphoinositide 3-kinase; and AIDS=Acquired ImmunoDeficiency Syndrome.

Abbreviations used herein have their conventional meaning within thechemical and biological arts, unless otherwise indicated.

The term “cell proliferation” refers to a phenomenon by which the cellnumber has changed as a result of division. This term also encompassescell growth by which the cell morphology has changed (e.g., increased insize) consistent with a proliferative signal.

The term “co-administration,” “administered in combination with,” andtheir grammatical equivalents, as used herein, encompassesadministration of two or more agents to an animal so that both agentsand/or their metabolites are present in the animal at the same time.Co-administration includes simultaneous administration in separatecompositions, administration at different times in separatecompositions, or administration in a composition in which both agentsare present.

The term “effective amount” or “therapeutically effective amount” refersto that amount of a compound described herein that is sufficient toeffect the intended application including, but not limited to, diseasetreatment. The therapeutically effective amount may vary depending uponthe intended application (in vitro or in vivo), or the subject anddisease condition being treated, e.g., the weight and age of thesubject, the severity of the disease condition, the manner ofadministration and the like, which can readily be determined by one ofordinary skill in the art. The term also applies to a dose that willinduce a particular response in target cells, e.g., reduction ofplatelet adhesion and/or cell migration. The specific dose will varydepending on the particular compounds chosen, the dosing regimen to befollowed, whether it is administered in combination with othercompounds, timing of administration, the tissue to which it isadministered, and the physical delivery system in which it is carried.

As used herein, the terms “treatment” and “treating” refer to anapproach for obtaining beneficial or desired results including, but notlimited to, therapeutic benefit and/or a prophylactic benefit. Bytherapeutic benefit is meant eradication or amelioration of theunderlying disorder being treated. Also, a therapeutic benefit isachieved with the eradication or amelioration of one or more of thephysiological symptoms associated with the underlying disorder such thatan improvement is observed in the patient, notwithstanding that thepatient may still be afflicted with the underlying disorder. Forprophylactic benefit, the compositions may be administered to a patientat risk of developing a particular disease, or to a patient reportingone or more of the physiological symptoms of a disease, even though adiagnosis of this disease may not have been made.

A “therapeutic effect,” as that term is used herein encompasses atherapeutic benefit and/or a prophylactic benefit as described above. Aprophylactic effect includes delaying or eliminating the appearance of adisease or condition, delaying or eliminating the onset of symptoms of adisease or condition, slowing, halting, or reversing the progression ofa disease or condition, or any combination thereof.

The term “subject” or “patient” refers to an animal, such as a mammal,for example a human. The methods described herein can be useful in bothhuman therapeutics and veterinary applications. In some embodiments, thepatient is a mammal, and in some embodiments, the patient is human. Forveterinary purposes, the term “subject” and “patient” include, but arenot limited to, farm animals including cows, sheep, pigs, horses, andgoats; companion animals such as dogs and cats; exotic and/or zooanimals; laboratory animals including mice, rats, rabbits, guinea pigs,and hamsters; and poultry such as chickens, turkeys, ducks, and geese.

“Radiation therapy” refers to exposing a patient, using methods andcompositions known to the practitioner, to radiation emitters such asalpha-particle emitting radionuclides (e.g., actinium and thoriumradionuclides), low linear energy transfer (LET) radiation emitters(i.e. beta emitters), conversion electron emitters (e.g. strontium-89and samarium-153-EDTMP), or high-energy radiation, including withoutlimitation x-rays, gamma rays, and neutrons.

“Signal transduction” is a process during which stimulatory orinhibitory signals are transmitted into and within a cell to elicit anintracellular response. A modulator of a signal transduction pathwayrefers to a compound which modulates the activity of one or morecellular proteins mapped to the same specific signal transductionpathway. A modulator may augment (agonist) or suppress (antagonist) theactivity of a signaling molecule.

The term “selective inhibition” or “selectively inhibit” as applied to abiologically active agent refers to the agent's ability to selectivelyreduce the target signaling activity as compared to off-target signalingactivity, via direct or indirect interaction with the target.

As used herein, the terms “PI3-kinase delta selective inhibitor” and“PI3-kinase δ selective inhibitor” generally refer to a compound thatinhibits the activity of the PI3-kinase δ isozyme more effectively thanother isozymes of the PI3K family (alpha, beta, and gamma). Forinstance, the PI3-kinase δ selective inhibitor may refer to a compoundthat exhibits a 50% inhibitory concentration (IC50) with respect to thedelta type I PI3-kinase that is at least 10-fold, at least 20-fold, atleast 50-fold, at least 100-fold, or lower, than the inhibitor's IC50with respect to the rest of the other type I PI3-kinases alpha, beta,and gamma).

Inhibition of PI3-kinase δ may be of therapeutic benefit in treatment ofvarious conditions, e.g., conditions characterized by an inflammatoryresponse including but not limited to autoimmune diseases, allergicdiseases, and arthritic diseases. Importantly, inhibition of PI3-kinaseδ function does not appear to affect biological functions such asviability and fertility.

“Inflammatory response” as used herein is characterized by redness,heat, swelling and pain (i.e., inflammation) and typically involvestissue injury or destruction. An inflammatory response is usually alocalized, protective response elicited by injury or destruction oftissues, which serves to destroy, dilute or wall off (sequester) boththe injurious agent and the injured tissue. Inflammatory responses arenotably associated with the influx of leukocytes and/or leukocyte (e.g.,neutrophil) chemotaxis. Inflammatory responses may result from infectionwith pathogenic organisms and viruses, noninfectious means such astrauma or reperfusion following myocardial infarction or stroke, immuneresponses to foreign antigens, and autoimmune diseases. Inflammatoryresponses amenable to treatment with the methods and compounds accordingto the invention encompass conditions associated with reactions of thespecific defense system as well as conditions associated with reactionsof the non-specific defense system.

The therapeutic methods of the invention include methods for thetreatment of conditions associated with inflammatory cell activation.“Inflammatory cell activation” refers to the induction by a stimulus(including, but not limited to, cytokines, antigens or auto-antibodies)of a proliferative cellular response, the production of solublemediators (including but not limited to cytokines, oxygen radicals,enzymes, prostanoids, or vasoactive amines), or cell surface expressionof new or increased numbers of mediators (including, but not limited to,major histocompatibility antigens or cell adhesion molecules) ininflammatory cells (including, but not limited to, monocytes,macrophages, T lymphocytes, B lymphocytes, granulocytes(polymorphonuclear leukocytes including neutrophils, basophils, andeosinophils) mast cells, dendritic cells, Langerhans cells, andendothelial cells). It will be appreciated by persons skilled in the artthat the activation of one or a combination of these phenotypes in thesecells can contribute to the initiation, perpetuation, or exacerbation ofan inflammatory condition.

“Autoimmune disease” as used herein refers to any group of disorders inwhich tissue injury is associated with humoral or cell-mediatedresponses to the body's own constituents.

“Transplant rejection” as used herein refers to an immune responsedirected against grafted tissue (including organs or cells (e.g., bonemarrow), characterized by a loss of function of the grafted andsurrounding tissues, pain, swelling, leukocytosis, andthrombocytopenia).

“Allergic disease” as used herein refers to any symptoms, tissue damage,or loss of tissue function resulting from allergy.

“Arthritic disease” as used herein refers to any disease that ischaracterized by inflammatory lesions of the joints attributable to avariety of etiologies.

“Dermatitis” as used herein refers to any of a large family of diseasesof the skin that are characterized by inflammation of the skinattributable to a variety of etiologies.

The term “particle size distribution” of a powder, or granular material,or particles dispersed in fluid, as used herein, is a list of values ora mathematical function that defines the relative amounts of particlespresent, sorted according to size. The d(0.1), d(0.5) and d(0.9) valuesindicate that 10%, 50% and 90% of the particles measured were less thanor equal to the size stated. For example, values of d(0.1)=3, d(0.5)=10and d(0.9)=100 mean that 10% of the particles were less than or equal to3 μm, 50% were less than or equal to 10 μm and 90% were less than orequal to 100 μm.

The terms d(0.1), d(0.5), and d(0.9) refer to the mesh size of a singlenotional sieve allowing 10%, 50%, or 90% of the total of all particlesof the sample to pass. Thus d(0.1)=2-100 μm indicates that the upperlimit of the particle size range defining the 10% of the smallestparticles in the sample is between 2 μm to 100 μm. Thus 10% of the totalparticles have a particle size of not more than the d(0.1) value meaningin this case that they have a maximum size of 2 μm to 100 μm.

Pharmaceutical Compositions

The present invention provides a pharmaceutical composition comprising aPTSA salt of TGR-1202 as described herein and one or morepharmaceutically acceptable excipients. For example, the PTSA salt ofTGR-1202 may have the particle size distribution described herein and/orcrystalline properties described herein.

In one aspect, the present invention provides a pharmaceuticalcomposition comprising crystalline Form B of PTSA salt of TGR-1202according to any embodiment herein (e.g., a crystalline Form B of PTSAsalt of TGR-1202 having a particle size distribution as defined in anyembodiment herein) and one or more pharmaceutically acceptableexcipients. In one embodiment, the pharmaceutical composition includes atherapeutically effective amount of crystalline Form B of PTSA salt ofTGR-1202 according to any embodiment herein (e.g., a crystalline Form Bof PTSA salt of TGR-1202 having a particle size distribution as definedin any embodiment herein). The pharmaceutical composition may includeone or more additional active ingredients as described herein.

The excipients may be selected from diluents, fillers, salts,disintegrants, binders, lubricants, glidants, dispersing agents,suspending agents, wetting agents, controlled release matrices,colorants, flavorings, buffers, stabilizers, solubilizers, andcombinations thereof.

The pharmaceutical compositions of the present invention can beadministered alone or in combination with one or more other activeagents. Where desired, the subject compounds and other agent(s) may bemixed into a preparation or both components may be formulated intoseparate preparations to use them in combination separately or at thesame time.

The PTSA salts of TGR-1202 and pharmaceutical compositions describedherein can be administered by any route that enables delivery of theTGR-1202 to the site of action, such as orally, bucally, intranasally,topically (e.g., transdermally), intraduodenally, parenterally(including intravenously, intraarterially, intramuscularally,intravascularally, intraperitoneally or by injection or infusion),intradermally, by intramammary, intrathecally, intraocularly,retrobulbarly, intrapulmonary (e.g., aerosolized drugs) orsubcutaneously (including depot administration for long term releasee.g., embedded-under the-splenic capsule, brain, or in the cornea),sublingually, anally, rectally, vaginally, or by surgical implantation(e.g., embedded under the splenic capsule, brain, or in the cornea).

The compositions can be administered in solid, semi-solid, liquid orgaseous form, or may be in dried powder, such as lyophilized form. Thepharmaceutical compositions can be packaged in forms convenient fordelivery, including, for example, solid dosage forms such as capsules,sachets, cachets, gelatins, papers, tablets, suppositories, pellets,pills, troches, and lozenges. The type of packaging will generallydepend on the desired route of administration. Implantable sustainedrelease formulations are also contemplated, as are transdermalformulations.

The amount of the PTSA salt of TGR-1202 to be administered is dependenton the mammal being treated, the severity of the disorder or condition,the rate of administration, the disposition of the compound and thediscretion of the prescribing physician. However, an effective dosage isin the range of about 0.001 to about 100 mg per kg body weight per day,preferably about 1 to about 35 mg/kg/day, in single or divided doses.For a 70 kg human, this would amount to about 0.05 to 7 g/day,preferably about 0.05 to about 2.5 g/day. An effective amount of thePTSA salt of TGR-1202 described herein may be administered in eithersingle or multiple doses (e.g., twice or three times a day).

The PTSA salt of TGR-1202 described herein may be used in combinationwith one or more of anti-cancer agents (e.g., chemotherapeutic agents),therapeutic antibodies, and radiation treatment.

The PTSA salt of TGR-1202 described herein may be formulated oradministered in conjunction with other agents that act to relieve thesymptoms of inflammatory conditions such as encephalomyelitis, asthma,and the other diseases described herein. These agents includenon-steroidal anti-inflammatory drugs (NSAIDs).

Preparations of various pharmaceutical compositions are known in theart. See, e.g., Anderson, Philip O.; Knoben, James E.; Troutman, WilliamG, eds., Handbook of Clinical Drug Data, Tenth Edition, McGraw-Hill,2002; Pratt and Taylor, eds., Principles of Drug Action, Third Edition,Churchill Livingston, N.Y., 1990; Katzung, ed., Basic and ClinicalPharmacology, Ninth Edition, McGraw Hill, 2003; Goodman and Gilman,eds., The Pharmacological Basis of Therapeutics, Tenth Edition, McGrawHill, 2001; Remingtons Pharmaceutical Sciences, 20th Ed., LippincottWilliams & Wilkins, 2000; Martindale, The Extra Pharmacopoeia,Thirty-Second Edition (The Pharmaceutical Press, London, 1999), all ofwhich are incorporated by reference herein in their entirety.

In one embodiment, the PTSA salt of TGR-1202 described herein isadministered at a dose selected to produce a concentration of compoundin the blood between about 20 to 5,000 ng/mL, and maintaining suchconcentration during a period of about 6 to 24 hours followingadministration. In another particular embodiment, the dose size andfrequency are selected to achieve a concentration of compound in theblood that is between about 50 to 2,500 ng/mL and maintain thatconcentration during a period of about 6 to 24 hours from the time ofadministration. In some embodiments, the dose size and frequency areselected to achieve a concentration of TGR-1202 in the blood that isbetween about 100 to 1,500 ng/mL following administration. In someembodiments, the dose size and frequency are selected to achieve aconcentration of TGR-1202 in the blood that is between about 100 to 750ng/mL over a period of about 6 to 24 hours from the time ofadministration. In further embodiments, the dose size and frequency isselected to achieve a C_(max), plasma level of PTSA salt of TGR-1202that is at least about 300 ng/mL and does not exceed about 10,000 ng/mL.

Methods of Treatment

The invention also provides methods of using the compounds andpharmaceutical compositions of the present invention to treat diseaseconditions, including, but not limited to, diseases associated withmalfunctioning of one or more types of PI3 kinase.

A detailed description of conditions and disorders mediated by PI3 δkinase activity is described in, for example, International PublicationNos. WO 2001/81346, WO 2010/123931, WO 2010/0111432 and WO 2010/057048,and in U.S. Publication No. US 2005/043239, each of which isincorporated herein by reference in its entirety for all purposes.

The treatment methods provided herein comprise administering to thesubject a therapeutically effective amount of a compound of theinvention. In one embodiment, the present invention provides a method oftreating an inflammation disorder, including autoimmune diseases in amammal. The method comprises administering to said mammal atherapeutically effective amount of a compound of the present invention.

It will be appreciated that the treatment methods of the invention areuseful in the fields of human medicine and veterinary medicine. Thus,the individual to be treated may be a mammal, preferably human, or otheranimals. For veterinary purposes, individuals include but are notlimited to farm animals including cows, sheep, pigs, horses, and goats;companion animals such as dogs and cats; exotic and/or zoo animals;laboratory animals including mice, rats, rabbits, guinea pigs, andhamsters; and poultry such as chickens, turkeys, ducks, and geese.

The invention also relates to a method of treating a hyperproliferativedisorder in a subject that comprises administering to said mammal atherapeutically effective amount of a compound of the present inventionor a pharmaceutically acceptable salt thereof. In some embodiments, saidmethod relates to the treatment of cancer such as acute myeloidleukemia, thymus, brain, lung, squamous cell, skin, eye, retinoblastoma,intraocular melanoma, oral cavity and oropharyngeal, bladder, gastric,stomach, pancreatic, bladder, breast, cervical, head, neck, renal,kidney, liver, ovarian, prostate, colorectal, esophageal, testicular,gynecological, thyroid, CNS, PNS, AIDS-related (e.g. Lymphoma andKaposi's Sarcoma) or viral-induced cancer. In some embodiments, saidmethod relates to the treatment of a non-cancerous hyperproliferativedisorder such as benign hyperplasia of the skin (e. g., psoriasis),restenosis, or prostate (e.g., benign prostatic hypertrophy (BPH)).

EXAMPLES

The examples and preparations provided below further illustrate andexemplify the methods of preparing compounds and compositions of theinvention. It is to be understood that the scope of the presentinvention is not limited in any way by the scope of the followingexamples and preparations.

XPRD patterns were acquired on a X′PertPRO MPD diffractometer equippedwith a Cu LFF X-ray tube (45 kV, 40 mA) using the following measurementparameters:

-   -   Scan mode: Continuous    -   PSD mode: Scanning    -   PSD Length: 2.12 o 2θ    -   Scan Range: 2.5 to 40 o2θ    -   Step size: 0.017/step    -   Scan Step time: 12.065 s    -   Divergence Slit type: Automatic    -   Incident Beam Monochromatic: No    -   Spinning: Yes

DSC spectra were acquired on a DSCQ2000 V24.11 Build 124.

Example 1 Preparation of the PTSA Salt of TGR-1202 (Form A)

7100 g of TGR-1202 was charged in a reactor containing 56.8 litres ofacetone and stirred at ambient temperature. 4680 g of p-toluenesulphonic acid was added and the reaction mixture was heated at atemperature of 60-65° C. for about 6 hours. The solvent was removed bydistillation under reduced pressure to obtain a wet residue. The wetresidue was degassed and allowed to cool to <20° C. Approximately 142litres of diethyl ether was then added and the resulting mixture wasstirred overnight, then filtered to obtain a solid mass which was washedwith diethyl ether and dried in vacuo to yield a solid mass. The solidmass was re-suspended in diethyl ether, stirred for 6 hours, and thenfiltered to yield a solid mass which was subsequently dissolved in 56.8litres of acetone, filtered through a HiFlow bed, and concentrated underreduced pressure. The resulting residue mass was stirred with waterovernight, then filtered and vacuum dried to yield 6600 g of the PTSAsalt of TGR-1202. HPLC: 99.21% and chiral purity of 99.64:0.36 (S:R).

Example 2 Preparation of the PTSA Salt of TGR-1202 (Form B)

1000 g of TGR-1202 was charged in a reactor containing 8 litres ofacetone and stirred at ambient temperature. 666 g of p-toluene sulphonicacid was then added and the reaction mixture was heated at a temperatureof 60-65° C. for about 6 hours. The solvent was removed by distillationunder reduced pressure to obtain a wet residue. The wet residue wasdegassed and allowed to cool to <20° C. Approximately 20 litres ofdiethyl ether was added and the resulting mixture was stirred overnight,then filtered to obtain a solid mass which was washed with diethyl etherand dried in vacuo to yield a solid mass which was then vacuum dried toyield 1150 g of the PTSA salt of TGR-1202. HPLC: 99.33% and chiralpurity: 99.61:0.39 (S:R).

Table 1 lists the XRPD pattern peaks and relative peak intensities forthe products of Examples 1 and 2.

TABLE 1 Example 1 Example 2 2 Theta Rel. Int. (%) 2 Theta Rel. Int. (%)5.0111 90.45 5.0368 99.91 6.1947 17.54 6.2245 23.13 7.3344 28.14 7.372227.98 7.8955 23.93 7.9120 15.31 8.4677 5.77 8.4862 3.10 10.0914 95.6610.1351 100.00 12.2407 38.58 12.2720 25.41 13.0311 18.40 12.4494 28.7413.3639 18.53 13.0607 18.06 15.9118 43.52 13.3947 6.18 16.1260 62.0515.9313 18.53 16.3442 43.31 16.1429 30.80 17.3580 33.21 16.3476 17.1517.7652 36.83 16.7829 3.73 18.1828 34.80 17.0690 9.84 19.0276 20.0417.3057 9.38 19.5461 38.96 17.4929 9.78 20.0004 50.44 17.7730 16.3320.8037 38.08 18.2136 15.19 21.1204 27.38 18.4160 12.24 21.7227 32.6018.7137 6.65 22.0621 78.29 19.1252 12.24 22.8416 33.03 19.6214 21.1023.1627 26.64 20.0040 25.77 24.4157 100.00 20.8028 15.49 25.3391 39.0421.1629 11.75 26.9321 23.79 21.4304 8.00 27.8349 14.56 21.7505 13.8928.4163 12.04 22.1114 47.85 29.2666 25.40 22.8531 9.68 30.3829 10.4023.1912 10.23 34.2390 3.40 24.4503 62.76 35.8360 4.32 25.0031 15.7838.0065 6.35 25.4128 18.58 38.9170 1.78 26.6621 4.52 26.9692 14.3527.4037 4.15 27.8432 9.39 28.5253 6.78 29.3252 17.35 29.6443 6.0830.4781 5.95 31.1600 1.72 32.5213 2.02 32.9371 2.57 34.1290 1.91 35.66803.36 36.0502 2.33 36.7594 2.46 37.1729 1.34 (Also see FIG. 1) (Also seeFIG. 2)

Table 1A lists the DSC endotherms for the PTSA salts prepared inExamples 1 and 2.

TABLE 1A Example 1 Example 2 ° C. ° C. 153.66 145.66

Example 3 Particle Size Analysis

Particle size (PSD) analysis of the PTSA salts of TGR-1202 prepared inExamples 1 and 2 was performed as described below using a Malvern MasterSizer 2000 with the Scirocco 2000 dry powder system. The instrumentparameters were:

-   -   Particle RI: 1.50    -   Absorption index: 0.1    -   Obscuration range: 1-5%    -   Analysis Model: General purpose    -   Sensitivity: Normal    -   Air Pressure: 2.0 Bar    -   Feed rate: 30%    -   Sample measurement Time: 6 Second    -   Background measurement Time: 6 Second    -   Number of measurement cycle: 1

Procedure: 5 g of TGR-1202 was transferred into a sample measuring cell(i.e., Scirocco 2000) and the airflow was started. Laser intensity wasmaintained more than 70%. Measurement was started and the vibration feedrate was increased in order to achieve the obscuration within thelimits. Once the obscuration came within the limit, measurement wasrecorded. Analysis was performed in triplicate and the results providedare an average of three experiments.

The particle size distribution (PSD) for the products of Examples 1 and2 is shown in Table 2.

TABLE 2 PTSA salt of TGR-1202 Example d(0.1) μm d(0.5) μm d(0.9) μmExample 1 8.01 147.41 510.73 Example 2 1.07 3.98 12.18

Example 4 Tableting of the PTSA Salts of TGR-1202

Tablet cores containing 200 mg (calculated as free base form) of thePTSA salts of TGR-1202 in Examples 1 and 2 were prepared in order tostudy their dissolution properties. These formulations are referred toas Examples 4a and 4b, respectively. The particle size distribution forthe PTSA snits of TGR-1202 prepared in Examples 1 and 2 is provided inExample 3.

Example 4a

Six different tablets were prepared (T1, T2, T3, T4, T5 and T6)containing the PTSA salt of TGR-1202 prepared in Example 1.

Example 4b

Six different tablets were prepared (T1, T2, T3, T4, T5 and T6)containing the PTSA salt of TGR-1202 prepared in Example 2.

Equipment Used:

The tabletting equipment used is shown in Table 3.

TABLE 3 Stage Equipment Name Manufacturing Sieves 20 or 24#, 40#, and60# sieve Rapid Mixer Granulator (RMG) Stirrer SS vessels FBD Multi Millwith 1.0 mm SS screen Conta Blender Compression Compression machineCadpress 17.8 × 8.8 mm, oval shaped D-Tooling Tablet de-dusters Metaldetectors (optional) Coating Coating machine Stirrer SS vesselsPackaging Heat Induction sealer

Preparation of Tablets:

The formulations of the tablets are shown in Table 4.

TABLE 4 Formulation 200 mg dose strength (calculated as free base form)S. No. Ingredients Example 4a Example 4b 1 PTSA salt of TGR-1202 260.20(salt) 260.20 (salt) 2 Hydroxypropyl betadex 270.00 270.00 3Microcrystalline cellulose 41.80 41.80 (Avicel PH101) 4 CroscarmelloseSodium 20.00 20.00 (Ac-Di-Sol) 5 Hydroxypropyl cellulose 6.00 6.00(Klucel LF) 6 Purified Water q.s q.s 7 Croscarmellose Sodium 70.00 70.00(Ac-Di-Sol) 8 Magnesium stearate 2.00 2.00 Core Tablet Weight 670.00670.00 Coating 9 Opadry II Brown 40L565004 — 20.10 10 Purified Water —q.s Coated Tablet Weight — 690.10

Manufacturing Process:

The tablets were prepared as follows:

-   -   1. Dispense the raw material quantities as required by the        formula.    -   2. Co-Sift the API with hydroxypropyl betadex, croscarmellose        sodium and microcrystalline cellulose through a #40 mesh and        collect in an in-process bulk container or double polyethylene        bags.    -   3. Dissolve the weighed quantity of hydroxypropyl cellulose        (Klucel LF) in purified water under stirring in a stainless        steel container to afford a clear solution (binder solution).    -   4. Load the blend of step 2 in a rapid mixer granulator (RMG)        and mix for 8 minutes at a slow impeller speed with the chopper        off.    -   5. Granulate the blend of step 4 with the binder solution from        step 3. Add additional water if required to obtain granules of a        suitable consistency. Perform additional kneading if required.        Perform intermittent scrapping, when required, during        granulation.    -   6. Dry the wet mass in fluid bed drier (FBD) at an inlet        temperature not more than 65° C. until the loss on drying (LOD)        is below 3.0% w/w. Perform intermittent scrapping, when        required.    -   7. Pass the dried granules of step 6 through a 24# sieve and        collect the sifted granules separately. Mill 24# retained        granules through a Multi mill equipped with a 1.0 mm stainless        steel screen at a fast speed with the knives forward. Collect        the milled granules and sift through a 24# sieve equipped on a        vibratory sifter. Collect the 24# pass granules in a separate        polybag.    -   8. If required, repeat step 7 once again for the 24# retained        granules. Collect in the in-process bulk container or double        polyethylene bags.    -   9. Sift croscarmellose sodium through a #40 and mix with the        granules of step in a Conta blender at 12 rpm for 8 minutes.    -   10. Sift magnesium stearate through a #60 mesh and mix with the        blend of step 9 in a Conta blender at 12 rpm for 3 minutes.    -   11. Unload the lubricated granules into double polythene bags.        Weigh and record the weight of the lubricated granules.    -   12. Compress the lubricated blend with 17.8×8.8 mm, oval shape        punches with an average weight of 670.00 mg/tab.

The coating procedure described below was performed only for the tabletsof Example 4b.

-   -   13. Coating Solution preparation: Transfer a weighed quantity of        purified water to a container equipped with a mechanical        stirrer. Disperse Opadry II Brown 40L565004 in the purified        water with continuous stirring and mix for 45 minutes. Filter        the resulting suspension through a 100# sieve or nylon cloth.    -   14. Load the compressed tablets in to a coating pan and continue        coating till the required weight gain (2.5-3.5%) is achieved.

The in-process checks (IPC) data for the tablets of Examples 4a and 4bare shown in Tables 5 and 6, respectively.

TABLE 5 Example 4a: IPC data 200 mg strength Parameter Nominal AverageRange Weight [mg] 670.00 ± 5% 669.70 664.8-678.6 Thickness [mm] — 5.315.27-5.35 Hardness [Kp]  12.0 ± 4.0 12.2 11.2-13.1 Disintegration [min]For record only 14-15 min Friability Not More Than 1% 0.018%

TABLE 6 Example 4b: IPC data 200 mg strength Parameter Nominal AverageRange Weight [mg] 670.00 ± 5% 671.10 650-683 Thickness [mm] — —5.33-5.42 Hardness [Kp]  12.0 ± 4.0 — 12.53-14.83 Disintegration [min]For record only 16-17 min Friability Not More Than 1% 0.04%

Tablets taken orally remain one of the most effective means of treatmentavailable. The effectiveness of such dosage forms relies on the drugdissolving in the fluids of the gastrointestinal tract prior toabsorption into the systemic circulation. The rate of dissolution of thetablet is therefore relevant to its performance.

Dissolution of cores from Examples 4a and 4b was performed using thefollowing parameters: 900 ml of a 0.1N HCl aqueous solution with 1%sodium lauryl sulfate (SLS), paddle apparatus according to USP apparatus1, paddle speed of 100 rounds per minute, and 6 runs (n=6). The criteriafor dissolution in media with a pH of 1 is not less than 75% (Q=75) in120 minutes. The dissolution results are shown in Table 7 below.

TABLE 7 Example 4a Example 4b Time (min) % Drug Release RSD % DrugRelease RSD 10 24 10.1 7 11.7 15 39 8.3 15 12.4 30 70 10.3 44 8.9 45 7812.3 72 5.6 60 79 13.3 91 2.3 90 81 12.8 98 2.4 120 82 12.1 100 2.6

Example 5 Pharmacokinetics

The oral bioavailability of tablets (Examples 4a and 4b) were evaluatedin human healthy volunteers. The protocol for the pharmacokineticsstudies is provided below.

All healthy subjects were fasted overnight (12 hours) before dosing andfasting was continued until 4.0 hours after administration of the testformulation. Test formulations (tablets) were prepared as provided inExamples 4a and 4b. Blood samples were collected, and placed into amicro centrifuge tube containing an anticoagulant. The blood sampleswere centrifuged immediately and separated plasma samples were frozen atbelow −80° C. and stored until analysis. The plasma concentrations oftest items in all samples were analyzed by LC-MS/MS. Pharmacokineticparameters (i.e., C_(max), AUC_(0-t), T_(max), and t_(1/2)) wereestimated.

The tablet composition comprising a PTSA salt of TGR-1202 preparedaccording to Example 2 exhibited a C_(max) about 2.5 fold and an areaunder the curve (AUC) about 1.9 fold greater than that of the tabletcomposition comprising a PTSA salt of TGR-1202 prepared according toExample 1. The results are provided in Table 8 below.

TABLE 8 Human Pharmacokinetics Parameters Tablets of Example 4a Tabletsof Example 4b # Healthy Volunteers 12 12 Dose (mg) 200 200 MedianT_(max) (hr) 2 3 C_(max) (ng/mL) 155.97 395.49 *T_(1/2) (hr) 74.92 73.59AUC_(0-t) (μg × hr/mL) 5.06 9.43 AUC_(0-inf) 7.28 13.92 (μg × hr/mL)

Although the invention herein has been described with reference toparticular embodiments, it is to be understood that these embodimentsare merely illustrative of the principles and applications of thepresent invention. It is therefore to be understood that numerousmodifications may be made to the illustrative embodiments and that otherarrangements may be devised without departing from the spirit and scopeof the present invention as described above. It is intended that theappended claims define the scope of the invention and that methods andstructures within the scope of these claims and their equivalents becovered thereby.

All publications, patents and patent applications cited in thisapplication are herein incorporated by reference to the same extent asif each individual publication, patent or patent application wasspecifically and individually indicated to be incorporated herein byreference.

The invention claimed is:
 1. A method for the treatment of non-Hodgkin'slymphoma, chronic lymphocytic leukemia, peripheral T-cell lymphoma, orHodgkin's lymphoma comprising the step of administering to a subject inneed thereof an effective amount of a p-toluenesulfonic acid salt of thecompound

wherein the salt has a d(0.9) of from about 5 to about 50 μm.
 2. Amethod for treating non-Hodgkin's lymphoma in a subject in need thereofcomprising the step of administering to the subject an effective amountof a p-toluenesulfonic acid salt of the compound

wherein the salt has a d(0.9) of from about 5 to about 50 μm.
 3. Amethod for treating chronic lymphocytic leukemia in a subject in needthereof comprising the step of administering to the subject an effectiveamount of a p-toluenesulfonic acid salt of the compound

wherein the salt has a d(0.9) of from about 5 to about 50 μm.
 4. Amethod for treating follicular lymphoma in a subject in need thereofcomprising the step of administering to the subject an effective amountof a p-toluenesulfonic acid salt of the compound

wherein the salt has a d(0.9) of from about 5 to about 50 μm.
 5. Themethod of claim 1, wherein the salt has a d(0.9) of from about 5 toabout 25 μm.
 6. The method of claim 2, wherein the salt has a d(0.9) offrom about 5 to about 25 μm.
 7. The method of claim 3, wherein the salthas a d(0.9) of from about 5 to about 25 μm.
 8. The method of claim 4,wherein the salt has a d(0.9) of from about 5 to about 25 μm.
 9. Themethod of claim 1, wherein the salt has a d(0.9) of from about 5 toabout 15 μm.
 10. The method of claim 2, wherein the salt has a d(0.9) offrom about 5 to about 15 μm.
 11. The method of claim 3, wherein the salthas a d(0.9) of from about 5 to about 15 μm.
 12. The method of claim 4,wherein the salt has a d(0.9) of from about 5 to about 15 μm.
 13. Themethod of claim 1, wherein the salt has a d(0.5) of from about 1 toabout 10 μm.
 14. The method of claim 2, wherein the salt has a d(0.5) offrom about 1 to about 10 μm.
 15. The method of claim 3, wherein the salthas a d(0.5) of from about 1 to about 10 μm.
 16. The method of claim 4,wherein the salt has a d(0.5) of from about 1 to about 10 μm.
 17. Themethod of claim 1, wherein the salt has a d(0.5) of from about 2 toabout 5 μm.
 18. The method of claim 2, wherein the salt has a d(0.5) offrom about 2 to about 5 μm.
 19. The method of claim 3, wherein the salthas a d(0.5) of from about 2 to about 5 μm.
 20. The method of claim 4,wherein the salt has a d(0.5) of from about 2 to about 5 μm.
 21. Themethod of claim 1, wherein the salt exhibits a XRPD pattern having oneor more peaks selected from 5.0, 10.1, 22.1, and 24.5±0.2° 2Θ.
 22. Themethod of claim 2, wherein the salt exhibits a XRPD pattern having oneor more peaks selected from 5.0, 10.1, 22.1, and 24.5±0.2° 2Θ.
 23. Themethod of claim 3, wherein the salt exhibits a XRPD pattern having oneor more peaks selected from 5.0, 10.1, 22.1, and 24.5±0.2° 2Θ.
 24. Themethod of claim 4, wherein the salt exhibits a XRPD pattern having oneor more peaks selected from 5.0, 10.1, 22.1, and 24.5±0.2° 2Θ.
 25. Themethod of claim 1, wherein the ratio of p-toluenesulfonic acid to thecompound

is about 1:1.
 26. The method of claim 2, wherein the ratio ofp-toluenesulfonic acid to the compound

is about 1:1.
 27. The method of claim 3, wherein the ratio ofp-toluenesulfonic acid to the compound

is about 1:1.
 28. The method of claim 4, wherein the ratio ofp-toluenesulfonic acid to the compound

is about 1:1.
 29. A method for the treatment of indolent non-Hodgkin'slymphoma comprising the step of administering to a subject in needthereof an effective amount of a p-toluenesulfonic acid salt of thecompound

wherein the salt has a d(0.9) of from about 5 to about 50 μm.
 30. Themethod of claim 29, wherein the salt has a d(0.9) of from about 5 toabout 25 μm.
 31. The method of claim 29, wherein the salt has a d(0.9)of from about 5 to about 15 μm.
 32. The method of claim 29, wherein thesalt has a d(0.5) of from about 1 to about 10 μm.
 33. The method ofclaim 29, wherein the salt has a d(0.5) of from about 2 to about 5 μm.34. The method of claim 29, wherein the salt exhibits a XRPD patternhaving one or more peaks selected from 5.0, 10.1, 22.1, and 24.5±0.2°2Θ.
 35. The method of claim 29, wherein the ratio of p-toluenesulfonicacid to the compound

is about 1:1.
 36. The method of claim 1, wherein the salt isadministered orally.
 37. The method of claim 2, wherein the salt isadministered orally.
 38. The method of claim 3, wherein the salt isadministered orally.
 39. The method of claim 4, wherein the salt isadministered orally.
 40. The method of claim 29, wherein the salt isadministered orally.